WO2018155525A1 - Anti-inflammatory agent - Google Patents

Abstract

The purpose of the present invention is to provide a novel anti-inflammatory agent. Provided is an anti-inflammatory agent including a liver hydrolysate as an active ingredient.

Description

Anti-inflammatory agent

The present invention relates to a highly safe anti-inflammatory agent.

With the super-aging society, the importance of efforts to prevent and treat lifestyle diseases such as metabolic diseases such as diabetes and obesity and cardiovascular diseases such as arteriosclerosis are increasing. In fact, an inhibitory action of IL-1β signal and NF-κB has been developed against diabetes. In chronic inflammatory diseases represented by rheumatoid arthritis, inflammatory cytokines such as TNF-α, IL-1, and IL-6 are abnormally produced and secreted, and these inflammatory cytokine inhibitors are excellent. Useful as an anti-inflammatory agent. For example, as a TNF-α inhibitor, an antibody drug such as infliximab is used as a therapeutic drug for rheumatoid arthritis (Non-patent Document 1). In addition, as the IL-6 inhibitor, the antibody drug tricizumab is used (Non-patent Document 2). Furthermore, protein preparations and antibody drugs such as anakinra, rilonacept, and canakinumab have been developed as IL-1 inhibitors (Non-patent Document 3).

However, most of the conventional inflammatory cytokine inhibitors are biological preparations typified by antibody drugs, and it has been reported that side effects such as abnormal liver function, infection, and anaphylaxis occur. Usage guidelines are created as follows.
Accordingly, an object of the present invention is to provide a novel anti-inflammatory agent based on the inhibition of inflammatory cytokines which is highly safe and can be widely used.

Therefore, the present inventor has examined the action on the production of inflammatory cytokines using various components. As a result, liver hydrolysates that have been widely used for improvement of liver function, improvement of hangover in chronic liver disease, etc. Since it has a strong IL-1 production inhibitory action, it has been found that it is useful as an anti-inflammatory agent by suppressing the production of inflammatory cytokines.

That is, the present invention provides the following [1] to [15].

[1] An anti-inflammatory agent comprising liver hydrolyzate as an active ingredient.
[2] An inflammatory cytokine production inhibitor comprising liver hydrolyzate as an active ingredient.
[3] An IL-1 production inhibitor comprising liver hydrolyzate as an active ingredient.
[4] A food composition for improving inflammation comprising liver hydrolyzate as an active ingredient.
[5] A food composition for suppressing inflammatory cytokine production comprising liver hydrolyzate as an active ingredient.
[6] A food composition for inhibiting IL-1 production comprising liver hydrolyzate as an active ingredient.
[7] Use of liver hydrolyzate for the production of an anti-inflammatory agent.
[8] Use of a liver hydrolyzate for producing an inflammatory cytokine production inhibitor.
[9] Use of a liver hydrolyzate for producing an IL-1 production inhibitor.
[10] A liver hydrolyzate for treating inflammatory diseases.
[11] A liver hydrolyzate for suppressing the production of inflammatory cytokines.
[12] A liver hydrolyzate for suppressing IL-1 production.
[13] A method for treating an inflammatory disease, comprising administering an effective amount of a liver hydrolyzate.
[14] A method for suppressing the production of inflammatory cytokines, comprising administering an effective amount of liver hydrolyzate.
[15] A method for inhibiting IL-1 production, comprising administering an effective amount of a hydrolyzate of liver.

Liver hydrolyzate has a strong inhibitory effect on IL-1 production, and therefore suppresses the production of inflammatory cytokines and is useful as an anti-inflammatory agent. In addition, liver hydrolyzate can be administered orally, has few side effects, and is excellent in safety and usability, so it can also be used as a food composition.

It is a figure which shows the effect | action with respect to IL-1 (beta) production of a liver hydrolyzate.

The active ingredient of the anti-inflammatory agent of the present invention is liver hydrolyzate. Liver hydrolyzate, also called liver hydrolyzate, liver extract, liver decomposed extract, or liver hydrolyzate, is obtained by hydrolyzing the liver with digestive enzymes etc., and is used as a liver function improving drug Is. As the liver, which is a raw material, fresh livers such as cows, pigs, bonito and whales are used. The obtained hydrolyzate is preferably used after being concentrated. Preferably, liver hydrolyzate defined as the pharmaceutical product is used.

Liver hydrolyzate contains various amino acids, nucleotides, vitamins, minerals, etc., mainly composed of low molecular weight peptides. More specifically, amino acids 19 to 78% by mass, peptides and proteins 17 to 73% by mass, sugars 1.8 to 11% by mass, lipids 0.005 to 0.04% by mass, and nucleic acids 0.7 to 2.5% by mass. %, Inorganic substances 1.6 to 5.4% by mass, vitamins 0.03 to 0.2% by mass, and glutathione 0.8% by mass or less are preferable. Also, amino acids 23 to 65% by mass, peptides and proteins 20 to 61% by mass, saccharides 2.2 to 8.6% by mass, lipids 0.006 to 0.035% by mass, nucleic acids 0.9 to 2.1% by mass , More preferably 1.9-4.5% by mass of inorganic substance, 0.04-0.15% by mass of vitamin, 0.7% by mass or less of glutathione, 29-52% by mass of amino acids, 25-49 of peptides and proteins % By weight, saccharides 2.8-6.9% by weight, lipids 0.008-0.03% by weight, nucleic acids 1.1-1.7% by weight, inorganics 2.4-3.6% by weight, vitamins More preferred are those containing 05 to 0.12% by mass and 0.6% by mass or less of glutathione.

Among these components, the amino acid composition is Ala 17 to 68 mg / g, Arg 0.6 to 4.4 mg / g, Asp 9 to 48 mg / g, Cystein 5 mg / g or less, Glu 18 to 63 mg / g, Gly 10-39 mg / g, His 3-17 mg / g, Ile 14-56 mg / g, Leu 26-98 mg / g, Lys 15-65 mg / g, Met 0.3-20 mg / g, Phe 13-46 mg / g, Pro 10-48 mg / g, Ser 12-49 mg / g, Thr 12-45 mg / g, Trp 3-13 mg / g, Tyr 1.6-41 mg / g, Val 18-71 mg / g are preferred. Also, Ala 21 to 57 mg / g, Arg 0.8 to 3.6 mg / g, Asp 11 to 40 mg / g, Cystein 4 mg / g or less, Glu 22 to 53 mg / g, Gly 13 to 32 mg / g, His 4 to 14 mg / g, Ile 17-47 mg / g, Leu 32-82 mg / g, Lys 18-54 mg / g, Met 0.4-17 mg / g, Phe 15-38 mg / g, Pro 12-40 mg / g, Ser 15 ~ 41 mg / g, Thr ~ 14-38 mg / g, Trp ~ 3.8-11 mg / g, Tyr ~ 1.9-34 mg / g, Val ~ 21-59 mg / g are more preferable, Ala ~ 26-45 mg / g, Arg ~ 1 2.9 mg / g, Asp 14-32 mg / g, Cystine 3 mg / g or less, Glu 27-42 mg / g, Gly 16-26 mg / g, His 5-11 mg / g, Ile 21-40 mg / g, Leu 40-66 mg / g, Lys 22-43 mg / g, Met 0.5-14 mg / g, Phe 19-31 mg / g, Pro 15-32 mg / g, Ser 18-33 mg / g, Thr 18 More preferred are 30 mg / g, Trp 4.8 to 8.4 mg / g, Tyr 2.4 to 27 mg / g, Val 27 to 48 mg / g.

In the present invention, a low molecular weight fraction of liver hydrolyzate can also be used. As the low molecular weight fraction of the liver hydrolyzate, a fraction having a molecular weight of 3000 or less, for example, a fraction obtained by collecting a fraction having a molecular weight of 3000 or less from the liver hydrolyzate using an ultrafiltration membrane or the like can be used.

As shown in Examples below, liver hydrolyzate and low molecular weight fraction thereof have an action of strongly suppressing the production of IL-1β, which is a kind of inflammatory cytokine, and an action of suppressing the expression of IL-1β mRNA. . Therefore, the liver hydrolyzate suppresses the production of inflammatory cytokines typified by IL-1, and is useful as a therapeutic agent for inflammatory diseases involving inflammatory cytokines and a food and drink composition for improving inflammation. Here, diseases involving inflammatory cytokines such as IL-1 include rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, sepsis, acute and chronic myelogenous leukemia, osteoporosis, lifestyle-related diseases and the like.

The anti-inflammatory agent of the present invention can be administered by oral administration, transdermal administration, enteral administration, intravenous administration, etc., and oral administration is more preferred. Examples of the preparation for oral administration include liquids, tablets, powders, fine granules, granules, capsules and the like, but liquids and tablets are preferable, and liquids are more preferable.

These preparations for oral administration include excipients such as lactose, mannitol, corn starch and crystalline cellulose, cellulose derivatives, binders such as gum arabic and gelatin, disintegrants such as carboxymethylcellulose calcium, talc, magnesium stearate And the like, solubilizers such as nonionic surfactants, flavoring agents, sweeteners, stabilizers, pH adjusters, water, ethanol, propylene glycol, glycerin and the like can be used. Further, a coating agent such as hydroxymethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, cellulose acetate phthalate, and methacrylate copolymer may be used.

Moreover, other active ingredients can also be mix | blended with the anti-inflammatory agent of this invention. Other active ingredients, vitamins B ₁ class; thiamine, thiamine mononitrate, thiamine hydrochloride, fursultiamine, bisbentiamine, Benhochiamin, thiamine disulfide, dicethiamine, thiamine propyl disulfide, and derivatives thereof, vitamin B ₂ compounds; riboflavin and derivatives and their salts, vitamin B ₃ compound; niacin, nicotinic acid, nicotinamide and derivatives and salts thereof, vitamin B ₅ like; panthenol, pantothenic acid and derivatives and salts thereof, vitamin B ₆ such; pyridoxine and derivatives and their salts, vitamin B ₁₂ compound; cyanocobalamin and derivatives and salts thereof, other vitamins vitamin a, vitamin C, vitamin E, vitamin K, vitamin P, dichloroacetate diisopropylamine, taurine, chondro Itine sulfate, royal jelly, caffeine, turmeric, Maria thistle, dandelion, dandelion, burdock, garlic, chrysanthemum, western yarrow, gardenia, sesame, radish carrot, asparagus, onion, chicory, medicinal salvia, Korean thistle (artichoke), Wolfberry, leguminous and iridaceae plants, pokeweed, elba de pasarinho, sete sangria, akamegashiwa, tea, resveratrol, catechins, berberine, rosemary, bean extract, metformin, bromelain, arginine, black cohosh, ibrite , Propolis, Samenankotsu, Nigahakka, Alfalfa, Astragalus, Belladonna, Yellow Thistle, Marigold, Chamomile, Devils Crow, Touki, Echinacea, Elder, Eucalyptus Oil, Tsushirogiku, ginger, ginseng, golden seal, Centella asiatica, hop, birch, lavender, licorice, saw palmetto, spirulina, St. John's wort, tea tree oil, valerian, wild yam, and the like.

Moreover, since the anti-inflammatory agent of the present invention is highly safe and can be administered orally, in addition to pharmaceuticals, quasi-drugs, health functional foods, sports drinks, rehabilitation drinks, functional foods such as pet foods, etc. It can also be used as a food composition.

The content of liver hydrolyzate in the anti-inflammatory agent of the present invention varies depending on the administration form, but is usually preferably 0.001 to 10% by mass, more preferably 0.001 to 5% by mass as a dry weight. Further, the daily dosage of liver hydrolyzate in the anti-inflammatory agent of the present invention is preferably 100 mg to 1076 mg, more preferably 383 mg to 623 mg, and further preferably 351 mg to 680 mg as a dry weight.

Next, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

Production Example 1
The filter part of an ultrafiltration membrane (MICROCON (registered trademark) Centrifugal Devices YM-3) capable of fractionation at a molecular weight of 3000 is prewashed with a phosphate buffer (PBS). A 100 mg / mL liver hydrolyzate (manufactured by Zeria Shinyaku Kogyo) stock is diluted to 12 mg / mL with PBS, and a part of the sample is stored for cell addition (sample before fractionation). Distribute 500 μL of diluted 12 mg / mL sample to ultrafiltration membrane, centrifuge at 12000 rpm, 4 ° C., 260 min, and collect filtrate fraction (liver hydrolyzate low molecular fraction: under Molecular Weight 3000: uMW3k) did.

Test example 1
The number of RAW264.7 cells was adjusted to 1.5 × 10 ⁶ cells / dish, seeded, and cultured overnight. Both the pre-fractionation sample or the filtrate fraction were diluted 20-fold with EMEM medium (final concentration about 0.6 mg / mL). As a control, PBS was diluted 20-fold with EMEM medium instead of the sample. Each sample (control, pre-fractionation, filtrate fraction) diluted with a medium was added to each dish group. After 24 hours of culture, lipopolysaccharide (LPS) was added to a final concentration of 1.0 μg / mL. The mixture was allowed to stand for 3 hours, RNA was collected, and cDNA was synthesized by reverse transcription. Next, EEF1A1 and IL-1β were measured by RT-RCR.

The result is shown in FIG.
In the RAW cells stimulated with LPS, the mRNA expression level of inflammation-related genes was compared between the liver hydrolyzate group and the Cont group to which PBS was added instead of the liver hydrolyzate. The inflammation suppression effect of (PLH: before fractionation) was suggested. Similarly, a significant decrease in IL-1β mRNA expression level was also observed in the liver hydrolyzate low molecular fraction (uMW3k) group.
From the above results, it was suggested that liver hydrolyzate has an anti-inflammatory effect, and a physiologically active substance exhibiting an anti-inflammatory effect is present in a low molecular fraction having a molecular weight of 3000 or less in the liver hydrolyzate. .

Claims (15)

1. An anti-inflammatory agent containing liver hydrolyzate as an active ingredient.
2. Inflammatory cytokine production inhibitor containing liver hydrolyzate as an active ingredient.
3. An IL-1 production inhibitor containing liver hydrolyzate as an active ingredient.
4. A food composition for improving inflammation containing liver hydrolyzate as an active ingredient.
5. Food composition for suppressing inflammatory cytokine production using liver hydrolyzate as an active ingredient.
6. A food composition for inhibiting IL-1 production comprising liver hydrolyzate as an active ingredient.
7. Use of liver hydrolyzate for the manufacture of anti-inflammatory agents.
8. Use of liver hydrolyzate for production of inflammatory cytokine production inhibitor.
9. Use of liver hydrolyzate for IL-1 production inhibitor production.
10. Liver hydrolyzate for treating inflammatory diseases.
11. Liver hydrolyzate to suppress the production of inflammatory cytokines.
12. Liver hydrolyzate to suppress IL-1 production.
13. A method for treating inflammatory diseases, comprising administering an effective amount of liver hydrolyzate.
14. A method for suppressing the production of inflammatory cytokines, comprising administering an effective amount of liver hydrolyzate.
15. A method for inhibiting IL-1 production, comprising administering an effective amount of liver hydrolyzate.

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